Composition Analysis of Animal Feed by HR ICP-OES

Composition Analysis of Animal Feed by HR ICP-OES Abstract The High-Resolution Array ICP-OES equipped with Standard-Kit was used for the analysis of Al, B, Ca, Co, Cr, Cu, Fe, I, K, Na, Mn, Mg, Mo, Ni, P, S, V and Zn in an animal feed sample. The sample was digested in 3 ml HNO 3 and 0.5 ml HCl by microwave (TOPWAVE ), filled up to a total volume of 20 ml, subsequently diluted by factor 50 to give a nominal matrix concentration of 0.5 g/l and measured with an aqueous standard calibration routine without matrix matching using 1000 mg/l multiple and single element standards from SIGMA ALDRICH. Method robustness was further tested from comparison of multiple lines for elements prone to spectral interferences (e.g. Cu, Ni, P, V and Zn) and a spike recovery test. Trace impurities and matrix-elements including Calcium, Iodine, Magnesium, Sodium and Potassium were determined from the same sample with excellent precision using Dual View PLUS. That is agreement of results, good recovery rates and RSD values about 1% were realized using interference-free emission lines for all elements. However, for Iodine prominent Phosphorous interferences on the I178.215 nm emission line were successfully removed by the CSI-Tool, which rules out overestimation of Iodine in Phosphorous-rich matrices. Challenge Interference-free Dual View PLUS method, reduction of detection limits Matrix Animal feed (HNO 3, HCl) Purpose Method development for QC lab Matrix-specific detection limits smaller than 1 µg/l (ppb) for all metals and 5 ppb for Iodine were achieved here, which highlights the unique sensitivity of the benefiting QC labs dealing with animal feed samples. 1/9

Materials and Methods Calibration Table 1: Concentration of calibration standards Element Unit Cal.0 Cal.1 Cal.2 Cal.3 Cal.4 Al mg/l 0 0.625 1.25 10 - B mg/l 0 0.25 0.5 4 - Ca, K mg/l 0 12.5 25 200 - Co, Cr, Mo, Ni, V mg/l 0 0.125 0.25 2 - Cu, Zn mg/l 0 0.1875 0.375 3 8 Fe mg/l 0 0.5625 1.125 9 - I µg/l 0 0.2 - - - Mg mg/l 0 2.5 5 40 - Mn mg/l 0 0.1875 0.375 3 5 Na mg/l 0 5 10 80 200 P mg/l 0 10 20 160 - S mg/l 0 3.125 6.25 50 - Calibration Curves 2/9

Method Parameters Table 2: Plasma configurations and set-up of the sample introduction system Parameter Specification Power 1200 W Plasma gas flow 12 L/min Auxillary gas flow 0.5 L/min Nebulizer gas flow 0.6 L/min Nebulizer Concentric nebulizer, 1.0 ml/min, borosilicate Spray chamber Cyclonic Spray Chamber, 50 ml, borosilicate Outer tube/inner tube Quartz / Quartz Injector Quartz, 2 mm Pump tubing PVC Sample pump rate 1.0 ml/min Rinse/ Read delay 45 s Pump fast run time 15 s Autosampler yes 3/9

Evaluation Parameters Table 3: Overview of method-specific evaluation parameters Element Line Plasma-view Integration- Read Evaluation [nm] Mode time [s] No. of pixel Baseline fit Polynomial degree Correction 2 Al 396.152 axial spectrum 3 3 ABC 1 auto - B 249.773 axial spectrum 3 3 ABC auto CSI 3 Ca 317.933 radial spectrum 3 3 ABC auto - Co 228.615 axial spectrum 3 3 ABC auto - Cr 267.716 axial spectrum 3 3 ABC auto - Cu 213.598 axial spectrum 3 3 ABC auto - 327.396 axial spectrum 3 3 ABC auto - Fe 238.204 axial spectrum 3 3 ABC auto - I 178.215 axial spectrum 3 3 ABC auto CSI, IEC 4 K 766.491 radial spectrum 3 3 ABC auto - Mg 285.213 axial PLUS spectrum 3 3 ABC auto - Mn 259.372 axial spectrum 3 3 ABC auto - Mo 202.030 axial spectrum 3 3 ABC auto - Na 589.592 radial PLUS peak 3 3 ABC auto - Ni 221.648 axial spectrum 3 3 ABC auto - 231.604 axial spectrum 3 3 ABC auto - 177.434 axial spectrum 3 3 ABC auto - P 178.224 axial spectrum 3 3 ABC auto - 213.618 axial spectrum 3 3 ABC auto - S 182.565 axial spectrum 3 3 ABC auto - V Zn 292.401 axial spectrum 3 3 ABC auto - 292.464 axial spectrum 3 3 ABC auto - 202.548 axial spectrum 3 3 ABC auto - 206.200 axial spectrum 3 3 ABC auto - 1 automatic baseline correction (ABC) 2 internal standard (IS) and/or mathematical corrections of spectral interferences 3 correction of iron interference by CSI-Tool recommended, but not used here 4 correction of phosphorous interference by CSI-Tool or inter-element correction (IEC) were used 4/9

Results and Discussion Table 4: Overview of results for animal feed sample Element Line [nm] 2 Potential Sample concentration in mg/l RSD Spike recovery test DL 3 interference 1 expected found % mg/l % µg/l Al 396.152 1.05 1.006 0.26 4 97.7 0.42 B 249.773 0.060 0.058 0.84 1.6 99.1 0.46 Ca 317.933 60.7 61.05±2.32 0.49 80 99.4-4 Co 228.615 < 0.01 < DL 5-0.8 97.3 0.39 Cr )6 267.716 P 0.016 0.004±0.0315 2.72 0.8 98.6 0.11 Cu )7 327.396 1.01 1.083±0.0818 0.33 3.2 97.6 0.52 213.598 P - 1.099±0.0664 0.38 3.2 98.4 1.54 Fe 238.204 3.21 3.163±0.072 0.18 3.6 97.7 0.13 I 178.215 P - 153.6 )8 0.27 8 98.8 4.97 K 766.491 56.2 55.1±0.5355 0.48 80 96.9-4 Mg 285.213 19.97 19.93±0.1207 0.71 16 98.7 0.16 Mn 259.372 0.70 0.698±0.0966 0.62 2 98 0.02 Mo 202.030 < 0.01 0.0031±0.0186 12.29 0.8 94.5 0.76 Na 589.592 14.88 16.24±4.623 0.31 80 103-4 Ni )9 221.648-0.0035±0.0406 5.87 0.8 103 0.50 231.604 OH 0.025 0.0032±0.0421 6.13 0.8 96.7 0.35 P )10 177.434-52.44±0.7913 0.24-4 - 4-4 178.224 I 52.69 52.88±0.9349 0.15 64 102 11.0 213.618 Cu - 49.22±0.686 0.41 64 96 10.5 S 180.672 19.22 20.53±0.1324 0.47 20 101 5.2 V 292.401 < 0.005 < DL - 0.8 102 0.23 292.464 - < DL - 0.8 102 0.2 Zn 202.548-21.66 )11 ±0.1445 0.6 3.2 102 0.19 206.200 22.9 21.58 )11 ±0.3608 0.36 3.2 95.7 0.25 1 likely spectral interferences on conventional ICP-OES instruments due to line overlap that are well-resolved (not present) on the 2 RSD from 3 replicate measurements per sample; not given when sample content was lower than detection limit 3 method-specific detection limit obtained from 3σ of SD on QC Blank (11 repetitive runs) 4 value not obtained 5/9

5 value below detection limit 6 expected value too high probably due to interference from P 267.711 nm line (seems plausible as Phosphorous concentration in the sample is very high) 7 excellent agreement of copper results for two lines shows that Cu 213.598 nm lines is not interfered by P 213.618 nm even for high Phosphorous contents 8 no confidence range due to number of calibration points; CSI-corrected value given here that was confirmed by inter-element correction (IEC, using P 177 line) 9 expected value too high probably due to interference from OH line; see good agreement of results for Ni 221 and Ni 231 on PlasmaQuant PQ 9000 Elite 10 agreement of Phosphorous results for three different lines two of which typically are spectrally interfered on conventional ICP-OES instruments 11 value exceeding the calibration range Application advantages Selection of High-Resolution spectra for animal feed sample (blue), Cal.1 (red) and blank (grey); automatic baseline fit (ABC, green) Fe (can be removed by CSI) Al 396.152 nm B 249.773 nm Ca 317.933 nm Co 228.615 nm 6/9

P Cu Cr 267.716 nm Cu 213.598 nm and P 213.618 nm Fe 238.204 nm K 766.491 nm Mg 285.213 nm Mn 259.372 nm Mo 202.030 nm Na 589.592 nm 7/9

Ni 231.604 nm S 180.672 nm V 292.401 nm V 292.464 nm Zn 206.200 nm 8/9

as-received HR ICP-OE spectrum I 178.215 nm Phosphorous Phosphorous that is a matrixelement does affect the Iodine trace detection (prominent P178 partly overlaps I178) Iodine - false positive results for Iodine, if this spectral interference from Phosphorous is not removed after interference removal by CSI-Tool I 178.215 nm when a Phosphorous spectrum is subtracted a wellresolved Iodine signal with a FWHM of 2.4 pm is obtained Reference: ICP_OES_24_16_en.docx This document is true and correct at the time of publication; the information within is subject to change. Other documents may supersede this document, including technical modifications and corrections. Content may be used without written permission but with citation of source. 2016 Publisher: 9/9